Program Director/Principal Investigator (Last, First, Middle): Nestler, Eric, J. Project Summary This R01 grant is aimed at better understanding the molecular mechanisms by which chronic exposure to drugs of abuse induces long-lasting changes in the brain's reward circuits that contribute to the complex behavioral abnormalities that define an addicted state. Our work focuses on the transcription factor, AFosB, which is induced in the nucleus accumbens (NAc) and other key brain reward regions in response to chronic administration of virtually all drugs of abuse. AFosB is unique in that it accumulates to appreciable levels only after chronic drug exposure and, because of its unusual stability, persists for weeks-months of withdrawal. AFosB is thus one mechanism by which chronic drug exposure can drive long-lasting changes in gene expression that contribute to addiction. Indeed, considerable evidence supports the view that induction of AFosB in NAc mediates a state of heightened reward and motivation that could contribute to aspects of the addiction process. In this requested R37 renewal, we will characterize the precise mechanisms through which AFosB exerts these actions. Using state-of-the-art genome-wide chromatin assays, we will identify the genes in the NAc that are direct targets for AFosB in the context of cocaine and opiate self-administration. Such genes, in turn, provide novel insight into the molecular and cellular basis of drug-induced neural and behavioral plasticity. Interestingly, when AFosB binds to its target genes, it can either activate or repress them. Our hypothesis is that such activation vs. repressive actions are determined by the chromatin milieu of the affected gene. As well, partially non-overlapping genes are regulated by AFosB in NAc in response to the two drugs. Although this is likely mediated in part by the partly distinct subsets of neurons in which the drugs induce AFosB, we once again hypothesize that this specificity is also mediated partly by differences in other chromatin changes that cocaine and opiates induce at specific genes. In addition to exploring AFosB's regulation of target genes, we will also characterize several mechanisms that are crucial in determining the amount and activity of AFosB induced in the NAc. Together, these studies will identify many novel actions of drugs of abuse which can be exploited for the development of improved diagnostic tests and treatments for addiction. PHS 398/2590 (Rev. 06/09) Page 2 Continuation Format Page ProgramDirector/PrincipalInvestigator(Last,First,Middle): Nestler,EriC,J. DETAILED BUDGET FOR INITIAL BUDGET PERIOD FROM THROUGH DIRECT COSTS ONLY 4/1/16 3/31/17 List PERSONNEL (Applicant organization only) Use Cal, Acad, or Summer to Enter Months Devoted to Project Enter Dollar Amounts Requested (omit cents) for Salary Requested and Fringe Benefits ROLE ON Cal. Acad. Summer INST.BASE SALARY FRINGE NAME PROJECT Mnths Mnths Mnths SALARY REQUESTED BENEFITS TOTAL Eric Nestler PD/PI 1.2 181,500 18,150 5,082 23,232 Li Shen Faculty 1.2 103,378 0 0 0 Deena Walker Postdoc 6 49,680 24,840 6,955 31,795 Jaclyn Rabkin Grad Student 12 33,500 33,500 0 33,500 Erin Calipari Postdoc 6 48,000 24,000 6,720 30,720 Dominika Burek Res. Asst 12 35,000 35,000 9,800 44,800 SUBTOTALS - CONSULTANT COSTS EQUIPMENT (Itemize) SUPPLIES (Itemize by category) General Supplies - 94,510 Animal Costs - 40,490 ChlP-seq and RNA-seq -18,000 TRAVEL Society for Neuroscience annual meeting INPATIENT CARE COSTS OUTPATIENT CARE COSTS ALTERATIONS AND RENOVATIONS (Itemize by category) OTHER EXPENSES (Itemize by category) Publication - 5,000 Bioinformatic Services - 25,000 Grad Student Tuition - 3,591; Health Insurance - 4,909 CONSORTIUM/CONTRACTUAL COSTS SUBTOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD CONSORTIUM/CONTRACTUAL COSTS TOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD PHS 398 (Rev. 08/12 Approved Through 8/31/2015) Page 3_ 135,490 28,557 164,047 153,000 3,000 38,500 DIRECT COSTS (/<em 7a, Face Page; $ 358,547 FACILITIES AND ADMINISTRATIVE COSTS $ 358,547 OMB No. 0925-0001 Form Page 4